As lesões fibro-ósseas da região maxilomandibular constituem um grupo heterogêneo de patologias que incluem lesões de desenvolvimento, reativas (displásicas) e neoplásicas, sendo que as duas principais entidades incluídas neste grupo são a displasia fibrosa e o fibroma ossificante. Devido a grande similaridade histológica entre estas patologias a avaliação das características clínicas e imaginológicas, juntamente com os aspectos histopatológicos, são o principal método de diagnóstico diferencial, porém devido a presença de diferentes níveis de maturidade destas patologias, em diversos casos os componentes histológicos assim como seu ordenamento são muito semelhantes. A busca por um processo de diagnóstico histológico mais fundamentado, hoje inexistente, é justificada pela diferença do curso clínico assim como pela eleição do tratamento entre a displasia fibrosa e o fibroma ossificante. Desta forma o propósito deste trabalho foi realizar uma análise retrospectiva descritiva das características clínico-demográficas e histopatológicas, e estabelecer um critério de diagnóstico diferencial que associe algumas características histopatológicas padronizadas juntamente com uma análise imunoistoquímica dos fatores de atividade de remodelação óssea destas lesões. Desta forma foram aplicados diferentes marcadores proteicos e moleculares em casos previamente diagnosticados de displasia fibrosa e fibroma ossificante, objetivando direcionar um critério de diagnóstico mais preciso. Nesse estudo retrospectivo foram avaliados 54 casos, sendo 30 casos de displasia fibrosa e 24 casos de fibroma ossificante, definidos após uma revisão nas análises das lâminas seguindo um padrão com algumas particularidades histomorfológicas para o diagnóstico diferencial. As características histológicas utilizadas para essa diferenciação, baseadas na revisão da literatura, foram: 1- limite entre a lesão e o tecido ósseo adjacente; 2- depósitos esféricos basofílicos (calcificações cementóides); 3- espaços negativos limítrofes entre o tecido ósseo e o tecido conjuntivo; 4- intensidade da celularidade do estroma e; 5- paralelismo das trabéculas ósseas lesionais. Após a reavaliação das lâminas seguindo a padronização de diferenciação com os cinco critérios histológicos mencionados anteriormente, foram reconsiderados os diagnósticos de 7 lâminas, ou seja 12,96% das amostras apresentaram alteração no diagnóstico anatomopatológico inicial. Posteriormente foram utilizados os seguintes imunomarcadores proteicos e moleculares do metabolismo ósseo em 9 lâminas de fibroma ossificante e em 7 lâminas de displasia fibrosa: interleucina 6 (IL-6), osteoprotegerina (OPG), osteocalcina (OCN) e o ligante do receptor do ativador do fator nuclear Kappa B (RANKL). A imunoexpressão destes marcadores foi observada nos seguintes locais: osteócitos, osteoblastos, osteoclastos e no estroma. Apenas a osteoprotegerina apresentou significância estatística nos osteócitos, osteoblastos e osteoclastos. A osteoprotegerina no estroma e os demais marcadores não apresentaram significância estatística em nenhum dos locais. / The fibro-osseous lesions of the maxillomandibular region constitute a heterogeneous group of pathologies that include developmental, reactive (dysplastic) and neoplastic lesions, and the two main entities included in this group are fibrous dysplasia and ossifying fibroma. Due to the great histological similarity between these pathologies the evaluation of the clinical and imaging characteristics, together with the histopathological aspects, are the main method of differential diagnosis, however due to the presence of different levels of maturity of these pathologies, in several cases the histological components as well as their arrangement are very similar. The search for a more substantiated histological diagnosis process, which does not exist today, is justified by the difference in the clinical course as well as by the choice of treatment between fibrous dysplasia and ossifying fibroma. Therefore, the purpose of this study was to perform a descriptive retrospective analysis of the clinical-demographic and histopathological characteristics and to establish a differential diagnosis criterion that associates some standard histopathological characteristics together with an immunohistochemical analysis of the bone remodeling activity factors of these lesions. In this way different molecular and protein markers were applied in previously diagnosed cases of fibrous dysplasia and ossifying fibroma, aiming at directing a more precise diagnosis criterion. In this retrospective study, 54 cases were evaluated, 30 cases of fibrous dysplasia and 24 cases of ossifying fibroma, defined after a review in the analysis of the slides following a pattern with some histomorphological peculiarities for the differential diagnosis. The histological characteristics used for this differentiation, based on the literature review, were: 1- border between the lesion and the adjacent bone tissue; 2- basophilic spherical deposits (cementoid calcifications); 3 - borderline negative spaces between the bone tissue and connective tissue; 4- intensity of stroma cellularity; 5- parallelism of lesionous trabeculae. After the reassessment of the slides following the standardization of differentiation with the five histological criteria mentioned above, the diagnoses of 7 slides were reconsidered, that is, 12.96% of the samples presented alterations in the initial anatomopathological diagnosis. Subsequently, the following protein and molecular biomarkers of bone metabolism were used on 9 ossifying fibrous slides and on 7 fibrous dysplasia slides: interleukin 6 (IL-6), osteoprotegerin (OPG), osteocalcin (OCN) and the activator receptor ligand Of nuclear factor Kappa B (RANKL). Immunoexpression of these markers was observed at the following sites: osteocytes, osteoblasts, osteoclasts and in the stroma. Only osteoprotegerin presented statistical significance in osteocytes, osteoblasts and osteoclasts. The osteoprotegerin in the stroma and the other markers did not present statistical significance in any of the sites.

Cystic fibrosis (CF) is one of the relatively common inherited diseases caused by a mutation in the gene encoding for CFTR protein, which forms a chloride channel that significantly affects ion homeostasis and the associated fluid management of the cell. This disease mainly affects the respiratory and digestive systems, being the most life-threatening in the respiratory tract. Patients with CF suffer from frequent and recurrent respiratory infections that lead to the development of chronic inflammation and gradual destruction of lung tissue. These lung infections, which are caused mostly by the opportunistic pathogen Pseudomonas aeruginosa, are the most common cause of morbidity and mortality in these patients. At present, antibiotics are used in the treatment of Pseudomonas aeruginosa infections, but new methods of antibacterial therapy need to be found to overcome the development of resistance. In addition to active immunization of CF patients against Pseudomonas aeruginosa, their passive immunization with specific chicken antibodies directed against this pathogen offers promising possibilities. This dissertation thesis is aimed to verify the prophylactic potential of hen IgY antibodies against the virulence factor Pseudomonas aeruginosa - lectin PAIIL, and to further develop an experimental...

Orientador: Gláucia Maria Ferreira da Silva Mazeto / Resumo: Introdução: A displasia fibrosa óssea (DFO) é uma doença rara, de origem genética somática, não hereditária, caracterizada por formação de tumores ósseos, resultando em dor óssea, deformidade e fraturas. Não há tratamento curativo e a cirurgia tem como finalidade a correção de fraturas e descompressão nervosa. A dor, principal sintoma desta doença, é fator limitante para as atividades diárias e prejudica a qualidade de vida. Para o alívio da dor óssea, os bisfosfonatos desempenham fundamental importância. Até o momento, não há consenso sobre o bisfosfonato mais adequado, o tempo ou o número ideal de ciclos de aplicação da medicação. Vários marcadores bioquímicos vêm sendo propostos no seguimento dos pacientes, ainda também sem definição sobre o mais adequado. A adequada quantificação de parâmetros relacionados à resposta terapêutica, particularmente da dor e da qualidade de vida, podem propiciar a monitorização objetiva e desempenhar papel norteador na abordagem destes pacientes. Objetivos: Avaliar a eficácia do pamidronato em pacientes com DFO, na melhora da dor óssea e da qualidade de vida. Casuística e métodos: Foram convidados a participar do estudo todos os pacientes portadores de DFO atendidos nos Ambulatórios de Endocrinologia, Ortopedia e Pediatria do Hospital das Clínicas da Faculdade de Medicina da Universidade Estadual Paulista “Júlio de Mesquita Filho” Unesp. Este estudo observacional, avaliou pacientes com DFO de forma prospectiva, por meio de coleta de dados dos... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Introduction: Fibrous Dysplasia (FD) is a rare disease of genetic origin, autosomal, not hereditary, characterized by the formation of bone tumors, resulting in bone pain, deformity and fracture. There is no curative treatment, and the only purpose of surgery is to correct fractures and nerve decompression. Pain is the main symptom, and limit daily activities impairing quality of life. For the relief of bone pain, bisphosphonates play a fundamental role. Until this moment, there is no consensus on which is the most adequate bisphosphonate to be used, the ideal timing of the application and the number of optimal medication application cycles. Several biochemical markers have been proposed in the follow-up of these patients, but yet there is no definition of the most appropriate. Proper quantification of parameters that evaluate therapeutics response, in particular pain and quality of life, can propitiate objective monitoring and play a guiding paper into approach these patients. Objectives: To evaluate the effect of using bisphosphonates in patients with FD, with perception of the improvement in relieving bone pain and quality of life. Methods: All the patients with FD attended in the Ambulatories of Endocrinology, Orthopedics and Pediatrics of the Hospital of the Medical School of Universidade Estadual Paulista “Júlio de Mesquita Filho” (Unesp) were invited to participate the research. This observational study, evaluate FD patients in a retrospective form, gathering medical r... (Complete abstract click electronic access below) / Mestre

Made available in DSpace on 2014-12-17T15:32:16Z (GMT). No. of bitstreams: 1 PedroPAS.pdf: 6126862 bytes, checksum: 3ae3ec2483d9e09d3a05fcd73782e862 (MD5) Previous issue date: 2008-02-27 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The giant cell fibroma is a benign neoplasm characterized by the presence of mono, bi or multinucleate cells, which can have a connection to the presence of mast cells. This research aims to analyze, descriptively and comparatively, the immunohystochemistry expression of the tryptase in mast cells of the giant cell f ibroma, f ibrous hyperplasia and samples of the normal oral mucosa. Thirty cases of giant cell fibroma, ten cases of fibrous hyperplasia and ten cases of normal oral mucosa were selected for the analysis of the immunohistochemistry expression, determination of the number of present mast cells, as well as their location and shape. It could be stated that there was a statistically signif icant difference (p<0,001) in relation to the quantity of mast cells among other samples analyzed where the giant cell f ibroma presented lesser quantity of mast cell and the hyperplasia showed higher concentration of this cellular type. Although the oral mucosa has presented a higher quantity of mast cells when compared to the giant cells fibroma, these were found in usual locations in the connective tissue in normal tissues. There could be noticed a statistically significant difference in relation to the number of non-granulated mast cells (p<0,001). On the areas of fibrosis, we could observe a statistically signif icant difference (p<0,006) among the samples. In relation to the present mast cells in perivascular location, no statistically signif icant difference was found. On the morphological analysis there was a predominance of oval mast cells. It was concluded that despite of the fact there was a lesser quantity of mast cells present in cases of giant cell f ibroma, they appeared to have a stronger relation to the present giant fibroblasts in this lesions, around 59,62%, being also evidenced a strong relation between these cells and the fibrosis areas in both cases of giant cell f ibroma and f ibrous hyperplasias and samples of normal oral mucosa, used as control group in our study, confirming, this way, the role of the mast cells as fibrinogenous inductor / O fibroma de c?lulas gigantes constitui-se de uma neoplasia benigna, caracterizada pela presen?a de c?lulas gigantes, mono, bi ou multinucleadas, c?lulas estas que podem guardar rela??o com a presen?a de mast?citos. O prop?sito desta pesquisa consistiu em analisar descritiva e comparativamente a express?o imuno-histoqu?mica da triptase em mast?citos de fibroma de c?lulas gigantes, hiperplasia fibrosa e esp?cimes de mucosa oral normal. Foram selecionados 30 casos de fibroma de c?lulas gigantes, 10 casos de hiperplasia fibrosa e 10 casos de mucosa oral normal, para a an?lise da express?o imuno-histoqu?mica, determina??o do n?mero de mast?citos presentes, bem como a sua forma e localiza??o. Constatou-se diferen?a estatisticamente significativa (p<0,001) em rela??o a quantidade de mast?citos entre os esp?cimes analisados, onde o fibroma de c?lulas gigantes apresentou a menor quantidade de mast?citos e a hiperplasia exibiu a maior concentra??o deste tipo celular. Embora a mucosa oral tenha apresentado uma maior quantidade de mast?citos quando comparado com os casos de f ibroma de c?lulas gigantes, estes se encontravam em localiza??es usuais no tecido conjuntivo em tecidos normais. Verif icou-se, diferen?a estatisticamente significativa, no que diz respeito ao n?mero de mast?citos n?o degranulados (p<0,001). Nas ?reas de fibrose, observamos diferen?a estatisticamente signif icativa (p<0,006) entre os esp?cimes. Com rela??o aos mast?citos presentes em localiza??o perivascular n?o se observou diferen?a estatisticamente significativa. Na an?lise morfol?gica verif icou-se uma predomin?ncia de mast?citos ovais. Concluiu-se que embora uma menor quantidade de mast?citos estivesse presente nos casos de fibroma de c?lulas gigantes, estes exibiam maior rela??o com os fibroblastos gigantes presentes nestas les?es em torno de 59,62%, sendo evidenciada tamb?m uma forte rela??o entre estas c?lulas e ?reas de fibrose tanto nos casos de fibroma de c?lulas gigantes como de hiperplasias fibrosas e esp?cimes de mucosa oral normal, utilizados como controle em nosso estudo, confirmando desta forma, o papel dos mast?citos como indutor fibrinog?nico

Fundação de Amparo a Pesquisa do Estado de Minas Gerais / The aim of this study was to evaluate the inflammatory response and repair process in subcutaneous tissue from the rats to four different sealers: Sealer 26, Epiphany, AH Plus and Endofill. Polyethylene tubes filled with fresh sealer were implanted in the four separate pockets created by blunt dissection in regions of the dorsum of each one of the third five male Wistar rats used. After 7, 14, 21, 42 and 60 days of post operatory, the implants were removed with the surrounding tissue, processed for inclusion in paraffin for the analysis histopatological. Qualitative and quantitative analysis was performed on HE sections as well as picro sirius (PS) sections. The intensity of the inflammatory response was evaluated and were scored as: 0- no inflammation;1- mild inflammation; 2- moderate inflammation or 3- severe inflammation. Fibrous capsule thickness was evaluated on PS sections using polarization. The measurements were subject to non parametric analysis. Results: In seven days the subcutaneous tissue of all animals showed characteristics corresponding to the severe inflammatory reaction (p> 0.05). At 14 and 21 days the response inflammatory tissued obtained with the Epiphany and AH Plus sealers showed statistically significant differences compared to the response of Endofill (p = 0.0005, p = 0.0003, respectively). This sealer remained the most aggressive at 42 (p = 0.0203) and 60 days (p = 0.0072) compared to Epiphany. There were no significant differences in the quantity of fibrous collagens among the sealers in the five periods. Conclusion: All tested sealers caused a severe inflammatory reaction in initial periods which decreased along the time. Endofill was the most aggressive sealer on all analyzed periods. The fibrous capsule was formed in all interfaces analyzed, presented himself as a protection developed by the tissue to the presence of an irritating material. / A presente pesquisa teve como objetivo verificar a resposta inflamatória e o processo de reparo em tecido subcutâneo de ratos a quatro cimentos endodônticos de composições distintas: Sealer 26®, Epiphany®, AH Plus® e Endofill. Neste estudo foram utilizados 35 ratos Wistar, sendo que em cada animal foram confeccionadas 4 lojas cirúrgicas subcutâneas na região dorsal. Em cada loja foi implantado um tubo de polietileno preenchido com um dos cimentos em teste. Após 7,14,21,42 e 60 dias de pós operatório, a região da pele em contato com o implante foi removida e os fragmentos foram processados para inclusão em parafina e posterior análise histopatológica. Metade dos cortes obtidos foram avaliados de acordo com a intensidade da resposta inflamatória atribuindo scores: 0= ausente,1= leve, 2=moderada e 3=severa que foram submetidos à análise estatística. A outra metade dos cortes foi corada em Picro-sirius Red para a análise da quantidade de fibras colágenas utilizando polarização ótica. Em sete dias, o tecido subcutâneo em todos os animais apresentou reação inflamatória severa, não sendo observadas diferenças estatísticas significantes entre os grupos (p>0.05). Aos 14 e 21 dias, os espécimes com os cimentos Epiphany® e AH Plus® mostraram diferenças estatísticas significantes quanto à intensidade da reação inflamatória quando comparados aos do cimento Endofill (p=0.0005, p=0.0003, respectivamente). Os espécimes com cimento Endofill mantiveram reação inflamatória intensa aos 42 (p=0.0203) e 60 dias (p=0.0072) comparado aos do cimento Epiphany®. Não foram encontradas diferenças quanto à quantidade de fibras colágenas na cápsula fibrosa entre os cimentos, em todos os tempos avaliados. Todos os cimentos testados provocaram uma reação inflamatória inicialmente severa que decresceu ao longo dos períodos avaliados. O Cimento Endofill se mostrou com maior potencial irritante em todos os períodos avaliados. A cápsula fibrosa, presente em todas as interfaces analisadas, se apresentou como uma proteção desenvolvida pelo tecido à presença do material irritante. / Mestre em Odontologia

Heart valve disease (HVD) or a damaged valve can severely compromise the heart's ability to pump efficiently. Balloon valvuloplasty is preferred on neonates with aortic valve stenosis. Even though this procedure decreases the gradient pressure across the aortic valve, restenosis is observed soon after balloon intervention. Tissue engineering heart valves (TEHV), using bone marrow stem cells (BMSCs) and biodegradable scaffolds, have been investigated as an alternative to current non-viable prosthesis. By observing the changes in hemodynamics following balloon aortic valvuloplasty, we could uncover a potential cause for rapid restenosis after balloon intervention. Subsequently, a tissue engineering treatment strategy based on BMSC mechanobiology could be defined. Understanding and identifying the mechanisms by which cytoskeletal changes may lead to cellular differentiation of a valvular phenotype is a first critical step in enhancing the promotion of a robust valvular phenotype from BMSCs.

Heart valve tissue engineering (HVTE) stands as a potential intervention that could reduce the prevalence of congenital heart valve disease in juvenile patients. Prior studies in our laboratory have utilized mechanobiological testing to quantify the forces involved in the development of heart valve tissue, utilizing a Flow-Stretch-Flexure (FSF) bioreactor to condition bone marrow stem cells (BMSCs)-derived valve tissue. Simulations have demonstrated that certain sets of flow conditions can introduce specific levels of oscillatory shear stress (OSS)-induced stimuli, augmenting the growth of engineered valves as well as influencing collagen formation, extracellular matrix (ECM) composition and gene expression. The computational findings discussed in this thesis outline the methods in which flow conditions, when physiologically relevant, induce specific oscillatory shear stresses which could not only lead to an optimized valve tissue phenotype (at 0.18≤ OSI≤ 0.23), but could identify native valve tissue remodeling indicative of aortic valve disease.

Calcific aortic valve disease (CAVD) is one of the most common causes of cardiovascular disease in North America. Mechanical factors have been closely linked to the pathogenesis of CAVD and may contribute to the disease by actively regulating the mechanobiology of valve interstitial cells (VICs). Mechanical forces affect VIC function through interactions between the VIC and the extracellular matrix (ECM). Studies have shown that the transfer of mechanical stimulus during cell-ECM interaction depends on the local material properties at hierarchical length scales encompassing tissue, cell and cytoskeleton. In this thesis, biomechanical tools were developed and applied to investigate hierarchical cell-ECM interactions, using VICs and valve tissue as a model system. Four topics of critical importance to understanding VIC-ECM interactions were studied: focal biomechanical material properties of aortic valve tissue; viscoelastic properties of VICs; transduction of mechanical deformation from the ECM to the cytoskeletal network; and the impact of altered cell-ECM interactions on VIC survival. To measure focal valve tissue properties, a micropipette aspiration (MA) method was implemented and validated. It was found that nonlinear elastic properties of the top layer of a multilayered biomaterial can be estimated by MA by using a pipette with a diameter smaller than the top layer thickness. Using this approach, it was shown that the effective stiffness of the fibrosa layer is greater than that of the ventricularis layer in intact aortic valve leaflets (p<0.01). To characterize the viscoelastic properties of VICs, an inverse FE method of single cell MA was developed and compared with the analytical half-space model. It was found that inherent differences in the half-space and FE models of single cell MA yield different cell viscoelastic material parameters. However, under particular experimental conditions, the parameters estimated by the half-space model are statistically indistinguishable from those predicted by the FE model. To study strain transduction from the ECM to cytoskeleton, an improved texture correlation algorithm and a uniaxial tension release device were developed. It was found that substrate strain fully transfers to the cytoskeletal network via focal adhesions in live VICs under large strain tension release. To study the effects of cell-ECM interactions on VIC survival, two mechanical stimulus systems that can simulate the separate effects of cell contraction and cell monolayer detachment were developed. It was found that cell sheet detachment and disrupted cell-ECM signaling is likely responsible for the apoptosis of VICs grown in culture on thin collagen matrices, leading to calcification. The studies presented in this thesis refine existing biomechanical tools and provide new experimental and analytical tools with which to study cell-ECM interactions. Their application resulted in an improved understanding of hierarchical valve biomechanics, mechanotransduction, and mechanobiology.

Calcific aortic valve disease (CAVD) is one of the most common causes of cardiovascular disease in North America. Mechanical factors have been closely linked to the pathogenesis of CAVD and may contribute to the disease by actively regulating the mechanobiology of valve interstitial cells (VICs). Mechanical forces affect VIC function through interactions between the VIC and the extracellular matrix (ECM). Studies have shown that the transfer of mechanical stimulus during cell-ECM interaction depends on the local material properties at hierarchical length scales encompassing tissue, cell and cytoskeleton. In this thesis, biomechanical tools were developed and applied to investigate hierarchical cell-ECM interactions, using VICs and valve tissue as a model system. Four topics of critical importance to understanding VIC-ECM interactions were studied: focal biomechanical material properties of aortic valve tissue; viscoelastic properties of VICs; transduction of mechanical deformation from the ECM to the cytoskeletal network; and the impact of altered cell-ECM interactions on VIC survival. To measure focal valve tissue properties, a micropipette aspiration (MA) method was implemented and validated. It was found that nonlinear elastic properties of the top layer of a multilayered biomaterial can be estimated by MA by using a pipette with a diameter smaller than the top layer thickness. Using this approach, it was shown that the effective stiffness of the fibrosa layer is greater than that of the ventricularis layer in intact aortic valve leaflets (p<0.01). To characterize the viscoelastic properties of VICs, an inverse FE method of single cell MA was developed and compared with the analytical half-space model. It was found that inherent differences in the half-space and FE models of single cell MA yield different cell viscoelastic material parameters. However, under particular experimental conditions, the parameters estimated by the half-space model are statistically indistinguishable from those predicted by the FE model. To study strain transduction from the ECM to cytoskeleton, an improved texture correlation algorithm and a uniaxial tension release device were developed. It was found that substrate strain fully transfers to the cytoskeletal network via focal adhesions in live VICs under large strain tension release. To study the effects of cell-ECM interactions on VIC survival, two mechanical stimulus systems that can simulate the separate effects of cell contraction and cell monolayer detachment were developed. It was found that cell sheet detachment and disrupted cell-ECM signaling is likely responsible for the apoptosis of VICs grown in culture on thin collagen matrices, leading to calcification. The studies presented in this thesis refine existing biomechanical tools and provide new experimental and analytical tools with which to study cell-ECM interactions. Their application resulted in an improved understanding of hierarchical valve biomechanics, mechanotransduction, and mechanobiology.